Publications

Despite serving the primary objective of ensuring that at least one sperm cell reaches and fertilizes an ovum, the male ejaculate (i.e. spermatozoa and seminal fluid) is a compositionally complex ‘trait’ that can respond phenotypically to subtle changes in conditions. In particular, recent research has shown that environmentally and genetically induced changes to ejaculates can have implications for offspring traits that are independent of the DNA sequence encoded into the sperm’s haploid genome. In this review, we compile evidence from several disciplines and numerous taxonomic systems to reveal the extent of such ejaculate-mediated paternal effects (EMPEs). We consider a number of environmental and genetic factors that have been shown to impact offspring phenotypes via ejaculates, and where possible we highlight the putative mechanistic pathways by which ejaculates can act as conduits for paternal effects. We also highlight how females themselves can influence EMPEs, and in some cases how maternally derived sources of variance may confound attempts to test for EMPEs. Finally, we consider a range of putative evolutionary implications of EMPEs, and suggest a number of potentially useful approaches for exploring these further. Overall our review confirms that EMPEs are both widespread and varied in their effects, although studies reporting their evolutionary effects are still in their infancy.

Mating often bears large costs to females, especially in species with high levels of sexual conflict over mating rates. Given the direct costs to females associated with multiple mating, which include reductions in lifespan and lifetime reproductive success, past research focused on identifying potential indirect benefits (through increases in offspring fitness) that females may accrue. Far less attention has, however, been devoted to understanding how costs of sexual interactions to females may extend across generations. Hence, little is known about the transgenerational implications of variation in mating rates, or the net consequences of maternal sexual activities across generations. Using the seed beetle, Callosobruchus maculatus, a model system for the study of sexual conflict, we investigate the effects of mating with multiple males versus a single male, and tease apart effects due to sexual harassment and those due to mating per se, over three generations. A multigenerational analysis indicated that females that were exposed to ongoing sexual harassment and who also were permitted to mate with multiple males showed no difference in net fitness compared to females that mated just once without ongoing harassment. Intriguingly, however, females that were continually harassed, but permitted to mate just once, suffered a severe decline in net fitness compared to females that were singly (not harassed) or multiply mated (harassed, but potentially gaining benefits via mating with multiple males). Overall, the enhanced fitness in multiply mated compared to harassed females may indicate that multiple mating confers transgenerational benefits. These benefits may counteract, but do not exceed (i.e., we found no difference between singly and multiply mated females), the large transgenerational costs of harassment. Our study highlights the importance of examining transgenerational effects from an inclusive (looking at both indirect benefits but also costs) perspective, and the need to investigate transgenerational effects across several generations if we are to fully understand the consequences of sexual interactions, sexual conflict evolution, and the interplay of sexual conflict and multi-generational costs and benefits.

Sex ratio allocation has important fitness consequences, and theory predicts that parents should adjust offspring sex ratio in cases where the fitness returns of producing male and female offspring vary. The ability of fathers to bias offspring sex ratios has traditionally been dismissed given the expectation of an equal proportion of X- and Y-chromosome-bearing sperm (CBS) in ejaculates due to segregation of sex chromosomes at meiosis. This expectation has been recently refuted. Here we used Peromyscus leucopus to demonstrate that sex ratio is explained by an exclusive effect of the father, and suggest a likely mechanism by which male-driven sex-ratio bias is attained. We identified a male sperm morphological marker that is associated with the mechanism leading to sex ratio bias; differences among males in the sperm nucleus area (a proxy for the sex chromosome that the sperm contains) explain 22% variation in litter sex ratio. We further show the role played by the sperm nucleus area as a mediator in the relationship between individual genetic variation and sex-ratio bias. Fathers with high levels of genetic variation had ejaculates with a higher proportion of sperm with small nuclei area. This, in turn, led to siring a higher proportion of sons (25% increase in sons per 0.1 decrease in the inbreeding coefficient). Our results reveal a plausible mechanism underlying unexplored male-driven sex-ratio biases. We also discuss why this pattern of paternal bias can be adaptive. This research puts to rest the idea that father contribution to sex ratio variation should be disregarded in vertebrates, and will stimulate research on evolutionary constraints to sex ratios—for example, whether fathers and mothers have divergent, coinciding, or neutral sex allocation interests. Finally, these results offer a potential explanation for those intriguing cases in which there are sex ratio biases, such as in humans.

Understanding and forecasting the effects of environmental change on wild populations requires knowledge on a critical question: do populations have the ability to evolve in response to that change? However, our knowledge on how evolution works in wild conditions under different environmental cir- cumstances is extremely limited. We investigated how environmental variation influences the evolutionary potential of phenotypic traits. We used published data to collect or calculate 135 estimates of evolvability of morpho- logical traits of European wild bird populations. We characterized the environmental favourability of each population throughout the species’ breed- ing distribution. Our results suggest that the evolutionary potential of morphological traits decreases as environmental favourability becomes high or low. Strong environmental selection pressures and high intra-specific competition may reduce species’ evolutionary potential in low- and high- favourability areas, respectively. This suggests that species may be least able to adapt to new climate conditions at their range margins and at the centre. Our results underscore the need to consider the evolutionary potential of populations when studying the drivers of species distributions, particularly when predicting the effects of environmental change. We discuss the utility of integrating evolutionary dynamics into a biogeographical perspective to understand how environmental variation shapes evolutionary patterns. This approach would also produce more reliable predictions about the effect of environmental change on population persistence and therefore on biodiversity.

Evans, J. P. & Garcia-Gonzalez, F.2016.The total opportunity for sexual selection and the integration of pre- and post-mating episodes of sexual selection in a complex world.Journal of Evolutionary Biology, 29: 2338-2361

It is well known that sexual selection can target reproductive traits during successive pre- and post-mating episodes of selection. A key focus of recent studies has been to understand and quantify how these episodes of sexual selection interact to determine overall variance in reproductive success. In this paper we review empirical developments in this field but also highlight the considerable variability in patterns of pre- and post-mating sexual selection, attributable to variation in patterns of resource acquisition and allocation, ecological and social factors, genotype-by-environment interaction, and possible methodological factors that might obscure such patterns. Our aim is to highlight how (co)variances in pre- and post-mating sexually selected traits can be sensitive to changes in a range of ecological and environmental variables. We argue that failure to capture this variation when quantifying the opportunity for sexual selection may lead to erroneous conclusions about the strength, direction or form of sexual selection operating on pre- and post-mating traits. Overall, we advocate for approaches that combine measures of pre- and post-mating selection across contrasting environmental or ecological gradients to better understand the dynamics of sexual selection in polyandrous species. We also discuss some directions for future research in this area.

Recognition of the ubiquity of female multiple mating has evoked an important shift in sexual selection research, emphasising the adaptive nature of female mating strategies. While phenotypic changes in female mating traits have been previously studied, little is known about the genetic basis of female mating behaviour and its potential to respond to selection at different stages throughout an individual’s life. Using a large quantitative genetic breeding design, we observed lifetime female mating behaviour in Drosophila melanogaster to examine the effect of female age and mating history on three key mating traits: courtship latency, mating latency and copula duration. Courtship latency (time until males initiate courtship) decreased with the cumulative number of females’ previous matings. Mating latency (defined here as the time between the beginning of courtship and the start of copulation) increased with female age, and copula duration was found to decrease as females aged. We calculated quantitative genetic estimates for mating traits in virgin females and at the females’ third mating to examine changes in the evolutionary potential of mating traits. We found considerable additive genetic variation in courtship latency and mating latency measured in virgin females. Copula duration displayed no heritable variation among females across sire families, but male effects were consistent with the idea that this trait is under male control. Heritability estimates differed significantly from zero in virgin females for courtship latency and mating latency. Heritability estimates did not differ significantly from zero when females were mating for the third time. However, overlapping 84% confidence intervals between heritability estimates obtained from virgin and mated females suggest that female mating strategies may have the potential to respond to selection at these different life stages.

Polyandry is widespread despite its costs. The sexually selected sperm hypotheses (‘sexy’ and ‘good’ sperm) posit that sperm competition plays a role in the evolution of polyandry. Two poorly studied assumptions of these hypotheses are the presence of additive genetic variance in polyandry and sperm competitiveness. Using a quantitative genetic breeding design in a natural population of Drosophila melanogaster, we first established the potential for polyandry to respond to selection. We then investigated whether polyandry can evolve through sexually selected sperm processes. We measured lifetime polyandry and offensive sperm competitiveness (P2) while controlling for sampling variance due to male x male x female interactions. We also measured additive genetic variance in egg-to-adult viability and controlled for its effect on P2 estimates. Female lifetime polyandry showed significant and substantial additive genetic variance and evolvability. In contrast, we found little genetic variance or evolvability in P2 or egg-to-adult viability. Additive genetic variance in polyandry highlights its potential to respond to selection. However, the low levels of genetic variance in sperm competitiveness suggest the evolution of polyandry may not be driven by sexy sperm or good sperm processes.

Females that mate with multiple males (polyandry) may reduce the risk that their eggs are fertilized by a single unsuitable male. About 25 years ago it was hypothesized that bet-hedging could function as a mechanism favoring the evolution of polyandry, but this idea is controversial because theory indicates that bet-hedging via polyandry can compensate the costs of mating only in small populations. Nevertheless, populations are often spatially structured, and even in the absence of spatial structure, mate choice opportunity can be limited to a few potential partners. We examined the effectiveness of bet-hedging in such situations with simulations carried out under two scenarios; (1) intrinsic male quality, with offspring survival determined by male phenotype (male’s ability to generate viable offspring), and (2) genetic incompatibility (offspring fitness determined non-additively by parental genotypes). We find higher fixation probabilities for a polyandrous strategy compared to a monandrous strategy if complete reproductive failure due to male effects or parental incompatibility is pervasive in the population. Our results also indicate that bet-hedging polyandry can delay the extinction of small demes. Our results underscore the potential for bet-hedging to provide benefits to polyandrous females and have valuable implications for conservation biology.

The trade-off between survival and reproduction is fundamental to life history theory. Sexual selection is expected to favour a live fast die young life history pattern in males due to increased risk of extrinsic mortality associated with obtaining mates. Sexual conflict may also drive a genetic trade-off between reproduction and lifespan in females. We found significant additive genetic variance in longevity independent of lifetime mating frequency, and in early life mating frequency. There was significant negative genetic covariance between these traits indicating that females from families characterized by high levels of multiple mating early in life die sooner than females that engage in less intense early life mating. Thus, despite heritable variation in both traits, their independent evolution is constrained by an evolutionary trade-off. Our findings indicate that, in addition to the well-known male-driven direct costs of mating on female lifespan (mediated by male harassment and the harmful effects of male seminal fluids), females with a genetic propensity to mate multiply live shorter lives. We discuss the potential role of sexual conflict in driving the evolutionary trade-off between reproduction and lifespan in Drosophila. More generally, our data show that, like males, females can exhibit a live fast die young life history strategy.

Garcia-Gonzalez, F., Yasui, Y. & Evans, J.P.2015.Risk-spreading by mating multiply is plausible and requires empirical attention.Proceedings of the Royal Society of London B, 282: 20150866

Garcia-Gonzalez, F. and Dowling, D. K.2015.Transgenerational effects of sexual interactions and sexual conflict: non-sires boost the fecundity of females in the following generation.Biology Letters, 11:20150067

The consequences of sexual interactions extend beyond the simple production of offspring. These interactions typically entail direct effects on female fitness, but may also impact the life histories of later generations. Evaluating the cross-generational effects of sexual interactions provides insights into the dynamics of sexual selection and conflict. Such studies can elucidate whether offspring fitness optima diverge across sexes upon heightened levels of sexual interaction among parents. Here, we found that, in Drosophila melanogaster, components of reproductive success in females, but not males, were contingent on the nature of sexual interactions experienced by their mothers. In particular, maternal sexual interactions with non-sires enhanced female fecundity in the following generation. This highlights the importance of non-sire influences of sexual interactions on the expression of offspring life histories.The consequences of sexual interactions extend beyond the simple production of offspring. These interactions typically entail direct effects on female fitness, but may also impact the life histories of later generations. Evaluating the cross-generational effects of sexual interactions provides insights into the dynamics of sexual selection and conflict. Such studies can elucidate whether offspring fitness optima diverge across sexes upon heightened levels of sexual interaction among parents. Here, we found that, in Drosophila melanogaster, components of reproductive success in females, but not males, were contingent on the nature of sexual interactions experienced by their mothers. In particular, maternal sexual interactions with non-sires enhanced female fecundity in the following generation. This highlights the importance of non-sire influences of sexual interactions on the expression of offspring life histories.

Theory assumes that postcopulatory sexual selection favours increased investment in testes size because greater numbers of sperm within the ejaculate increase the chance of success in sperm competition, and larger testes are able to produce more sperm. However, changes in the organization of the testes tissue may also affect sperm production rates. Indeed, recent comparative analyses suggest that sperm competition selects for greater proportions of sperm-producing tissue within the testes. Here, we explicitly test this hypothesis using the powerful technique of experimental evolution. We allowed house mice (Mus domesticus) to evolve via monogamy or polygamy in six replicate populations across 24 generations. We then used histology and image analysis to quantify the proportion of sperm-producing tissue (seminiferous tubules) within the testes of males. Our results show that males that had evolved with sperm competition had testes with a higher proportion of seminiferous tubules compared with males that had evolved under monogamy. Previously, it had been shown that males from the polygamous populations produced greater numbers of sperm in the absence of changes in testes size. We thus provide evidence that sperm competition selects for an increase in the density of sperm-producing tissue, and consequently increased testicular efficiency.

Polyandry (female multiple mating) has profound evolutionary and ecological implications. Despite considerable work devoted to understanding why females mate multiply, we currently lack convincing empirical evidence to explain the adaptive value of polyandry. Here we provide a direct test of the controversial idea that bet-hedging functions as a risk-spreading strategy that yields multi-generational fitness benefits to polyandrous females. Unfortunately, testing this hypothesis is far from trivial, and the empirical comparison of the across-generations fitness payoffs of a polyandrous (bet hedger) versus a monandrous (non-bet hedger) strategy has never been accomplished because of numerous experimental constraints presented by most ‘model’ species. In the present study we take advantage of the extraordinary tractability and versatility of a marine broadcast spawning invertebrate to overcome these challenges. We are able to simulate multi-generational (geometric mean) fitness among individual females assigned simultaneously to a polyandrous and monandrous mating strategy. Our approaches, which separate and account for the effects of sexual selection and pure bet-hedging scenarios, reveal that bet-hedging, in addition to sexual selection, can enhance evolutionary fitness in multiply-mated females. In addition to offering a tractable experimental approach for addressing bet-hedging theory, our study provides key insights into the evolutionary ecology of sexual interactions.

While model systems are useful in entomology, allowing generalizations to be made based on a few well-known species, they also have drawbacks. It can be difficult to know how far to generalize from information in a few species: are all flies like Drosophila? The use of model systems is particularly problematic in studying sexual selection, where variability among taxa is key to the evolution of different behaviors. A bias toward use of a few insect species, particularly Drosophila, is evident in the sexual selection and sexual conflict literature over the last several decades, although the diversity of study organisms has increased more recently. As the number of model systems used to study sexual conflict increased, support for the idea that sexual interactions resulted in harm to females decreased. Future work should choose model systems thoughtfully, combining well-known species with those that can add to the variation that allows us to make more meaningful generalizations.

In many species, females exposed to increased sexual activity experience reductions in longevity. Here, in Drosophila melanogaster, we report an additional effect on females brought about by sexual interactions; an effect that spans generations. We subjected females to a sexual treatment consisting of different levels of sexual activity, and then investigated patterns of mortality in their offspring. We found reduced probabilities of survival, increases in the rate-of-senescence, and a pattern of reduced mean longevities, for offspring produced by mothers that experienced higher levels of sexual interaction. We contend that these effects constitute trans-generational costs of sexual conflict – the existence or implications of which have rarely been considered previously. Our results indicate that ongoing exposure by mothers to male pre-copulatory interactions is itself sufficient to drive trans-generational effects on offspring mortality. Thus, we show that increases in maternal sexual activity can produce trans-generational effects that permeate through to latter life-stages in the offspring. This helps to elucidate the complex interplay between sex and ageing, and provides new insights into the dynamics of adaptation under sexual selection.

A key assumption underpinning major models of sexual selection is the expectation that male sexual attractiveness is heritable. Surprisingly, however, empirical tests of this assumption are relatively scarce. Here we use a paternal fullsib/half-sib breeding design to examine genetic and environmental variation in male mating latency (a proxy for sexual attractiveness) and copulation duration in a natural population of Drosophila melanogaster. As our experimental design also involved the manipulation of the social environment within each full-sibling family, we were able to further test for the presence of genotype-by-environment interactions (GEIs) in these traits, which have the potential to compromise mate choice for genetic benefits. Our experimental manipulation of the social environment revealed plastic expression of both traits; males exposed to a rival male during the sensitive period of adult sexual maturation exhibited shorter mating latencies and longer copulation durations than those who matured in isolation. However, we found no evidence for GEIs, and no significant additive genetic variation underlying these traits in either environment. These results undermine the notion that the evolution of female choice rests on covariance between female preference and male displays, an expectation that underpins indirect benefit models such as the good genes and sexy sons hypotheses. However, our results may also indicate depletion of genetic variance in these traits in the natural population studied, thus supporting the expectation that traits closely aligned with reproductive fitness can exhibit low levels of additive genetic variance.

In 1992 David Houle showed that measures of additive genetic variation standardized by the trait mean, CVA (the coefficient of additive genetic variation) and its square (IA), are suitable measures of evolvability. CVA has been used widely to compare patterns of genetic variation. However, the use of CVAs for comparative purposes relies critically on the correct calculation of this parameter. We reviewed a sample of quantitative genetic studies, focusing on sire models, and found that 45% of studies use incorrect methods for calculating CVA and that practices that render these coefficients meaningless are frequent. This may have important consequences for conclusions drawn from comparative studies. Our results are suggestive of a broader problem because miscalculation of the additive genetic variance from a sire model is prevalent among the studies sampled, implying that other important quantitative genetic parameters might also often be estimated incorrectly. We discuss the most prominent issues affecting the use of CVA and IA, including scale effects, data transformation, and the comparison of traits with different dimensions. Our aim is to increase awareness of the potential mistakes surrounding the calculation and use of evolvabilities, and to compile general guidelines for calculating, reporting and interpreting these useful measures in future studies.

Monogamy results in high genetic relatedness among offspring and thus it is generally assumed to be favored by kin selection. Female multiple mating (polyandry) has nevertheless evolved several times in the social Hymenoptera (ants, bees, and wasps), and a substantial amount of work has been conducted to understand its costs and benefits. Relatedness and inclusive fitness benefits are, however, not only influenced by queen mating frequency but also by paternity skew, which is a quantitative measure of paternity biases among the offspring of polyandrous females.We performed a large-scale phylogenetic analysis of paternity skew across polyandrous social Hymenoptera. We found a general and significant negative association between paternity frequency and paternity skew. High paternity skew, which increases relatedness among colony members and thus maximizes inclusive fitness gains, characterized species with low paternity frequency. However, species with highly polyandrous queens had low paternity skew, with paternity equalized among potential sires. Equal paternity shares among fathers are expected to maximize fitness benefits derived from genetic diversity among offspring. We discuss the potential for postcopulatory sexual selection to influence patterns of paternity in social insects, and suggest that sexual selection may have played a key, yet overlooked role in social evolution.

Garcia-Gonzalez, F.2012.Explainer: what is a null hypothesis?.The Conversation

In numerous species, egg chemoattractants play a critical role in guiding sperm towards unfertilized eggs (sperm chemotaxis). Until now the known functions of sperm chemotaxis include increasing the effective target size of eggs, thereby promoting sperm-egg encounters, and facilitating species recognition. Here we report that in the broadcast spawning mussel Mytilus galloprovincialis egg chemoattractants may play an unforeseen role in sexual selection by enabling sperm to effectively ‘choose’ between the eggs of different conspecific females. In an initial experiment we confirmed that sperm chemotaxis occurs in M. galloprovincialis by showing that sperm are attracted towards unfertilized eggs when given the choice of eggs or no eggs in a dichotomous chamber. We then conducted two cross-classified mating experiments, each comprising the same individual males and females crossed in identical male x female combinations but under experimental conditions that offered sperm ‘no-choice’ (each fertilization trial took place in a petri dish and involved a single male and female) or a ‘choice’ of a female’s eggs (sperm were placed in the centre of a dichotomous choice chamber and allowed to choose eggs from different females). We show that male-by-female interactions characterized fertilization rates in both experiments, and that there was remarkable consistency between patterns of sperm migration in the egg choice experiment and fertilization rates in the no-choice experiment. These results reveal that sperm differentially select eggs on the basis of chemical cues, thus exposing the potential for egg chemoattractants to mediate mate choice for genetically compatible partners. Given the prevalence of sperm chemotaxis across diverse taxa, our findings may have broad implications for sexual selection in other mating systems.

Male genitalia typically exhibit patterns of rapid and divergent evolution, and there is now considerable evidence that sexual selection is an important driver of these patterns of phenotypic variation. Female genitalia have been less well studied, and are generally thought to be relatively invariant. Here we use experimental evolution to show that sexual selection drives the correlated evolution of female and male genital morphology in the scarabaeine dung beetle Onthophagus taurus. Moreover, we use quantitative genetic analyses to provide a rare insight into the genetic architecture underlying morphological variation in female genital morphology, and uncover evidence of the genetic covariation with male genital morphology that is expected to arise under persistent sexual selection.

A key question in sexual selection is whether the ability of males to fertilize eggs under sperm competition exhibits heritable genetic variation. Addressing this question poses a significant problem, however, because a male’s ability to win fertilizations ultimately depends on the competitive ability of rival males. Attempts to partition genetic variance in sperm competitiveness, as estimated from measures of fertilization success, must therefore account for stochastic effects due to the random sampling of rival sperm competitors. In this contribution, we suggest a practical solution to this problem. We advocate the use of simple crossclassified breeding designs for partitioning sources of genetic variance in sperm competitiveness and fertilization success and show how these designs can be used to avoid stochastic effects due to the random sampling of rival sperm competitors. We illustrate the utility of these approaches by simulating various scenarios for estimating genetic parameters in sperm competiveness, and show that the probability of detecting additive genetic variance in this trait is restored when stochastic effects due to the random sampling of rival sperm competitors are controlled. Our findings have important implications for the study of the evolutionary maintenance of polyandry.

Whether species exhibit significant heritable variation in fitness is central for sexual selection. According to good genes models there must be genetic variation in males leading to variation in offspring fitness if females are to obtain genetic benefits from exercising mate preferences, or by mating multiply. However, sexual selection based on genetic benefits is controversial, and there is limited unambiguous support for the notion that choosy or polyandrous females can increase the chances of producing offspring with high viability. Here we examine the levels of additive genetic variance in two fitness components in the dung beetle Onthophagus taurus. We found significant sire effects on egg-to-adult viability and on son, but not daughter, survival to sexual maturity, as well as moderate coefficients of additive variance in these traits. Moreover, we do not find evidence for sexual antagonism influencing genetic variation for fitness. Our results are consistent with good genes sexual selection, and suggest that both pre- and postcopulatory mate choice, and male competition could provide indirect benefits to females.

Sexual selection is a potent force shaping multiple aspects of the interaction between the sexes, including the characters underlying reproductive success and sexual conflict, and may play an important role in determining the viability of populations. Experimental evolution is a methodological approach in which researchers either act as selective agents or establish the selective pressures operating on individuals to investigate changes in traits across generations and the genetic underpinning of these changes. Experimental evolution replicates the evolutionary process under controlled conditions and, by doing so, offers exceptional insights into the role of variation, selection and adaptation in evolution. Applied to the study of pre-copulatory (before mating) and postcopulatory (after mating) sexual selection, experimental evolution proves critical to understand the evolutionary consequences of male–malecompetition and femalemate choice, and the repercussions of concurrent or divergent interests between the sexes in regard to reproduction.

Sexual conflict facilitates the evolution of traits that increase the reproductive success of males at the expense of components of female fitness. Theory suggests that indirect benefits are unlikely to offset the direct costs to females from antagonistic male adaptations, but empirical studies examining the net fitness pay-offs of the interaction between the sexes are scarce. Here, we investigate whether matings with males that invest intrinsically more into accessory gland tissue undermine female lifetime reproductive success (LRS) in the cricket Teleogryllus oceanicus. We found that females incur a longevity cost of mating that is proportional to the partner’s absolute investment into the production of accessory gland products. However, male accessory gland weight positively influences embryo survival, and harmful ejaculate-induced effects are cancelled out when these are put in the context of female LRS. The direct costs of mating with males that sire offspring with higher viability are thus compensated by direct and possibly indirect genetic benefits in this species.

Selection imposed through sperm competition is commonly thought to promote the evolution of longer sperm, since sperm length is assumed to be positively associated with sperm swimming velocity. Yet, the basis for this assumption remains controversial, and there is surprisingly little intraspecific evidence demonstrating such a link between sperm form and function. Here, we show that sperm length and velocity are highly correlated in the sea urchin Heliocidaris erythrogramma, but importantly we report that failure to account for within-male variation in these sperm traits can obscure this relationship. These findings, in conjunction with the mounting evidence for extremely high levels of intraspecific variance in sperm traits, suggest that a functional link between sperm morphology and velocity may be more prevalent than what current evidence suggests. Our findings also suggest that selection for faster swimming sperm may promote the evolution of longer sperm, thereby supporting recent findings from macroevolutionary studies.

Male genital morphology is characterized by two striking and general patterns of morphological variation: rapid evolutionary divergence in shape and complexity, and relatively low scaling relationships with body size. These patterns of variation have been ascribed to the action of sexual selection. Among species, monogamous taxa tend to have relatively less complex male genital morphology than do polygamous taxa. However, although variation in male genital morphology can be associated with variation in mating and fertilization success, there is no direct evidence that sexual selection generates the evolutionary changes in male genital shape that underlie observed macroevolutionary patterns. Moreover, the hypothesis that sexual selection acts to reduce the scaling relationship between body and genital size is based entirely on the theoretical argument that male genitalia should be selected to provide an appropriate mechanical and/or stimulatory fit to the most commonly encountered female genitalia. Here, using the dung beetle Onthophagus taurus, we combine the power of experimental evolution with multivariate selection and quantitative genetic analyses to provide the most comprehensive evidence available of the form and evolutionary consequences of sexual selection acting on male genital morphology.

We report 16 polymorphic microsatellite loci from Heliocidaris erythrogramma, a common sea urchin endemic to temperate Australian waters. These microsatellites were tested in a minimum of 30 individuals, which yielded between five and 14 alleles per locus. Expected heterozygosity ranged from 0.52 to 0.92 with four loci deviating from Hardy–Weinberg expectations. These markers are expected to be useful for experimental studies involving paternity analysis and for quantifying population structure in H. erythrogramma across its geographic range.

Background: The determination of genetic variation in sperm competitive ability is fundamental to distinguish between post-copulatory sexual selection models based on good-genes vs compatible genes. The sexy-sperm and the good-sperm hypotheses for the evolution of polyandry require additive (intrinsic) effects of genes influencing sperm competitiveness, whereas the genetic incompatibility hypothesis invokes non-additive genetic effects. A male's sperm competitive ability is typically estimated from his fertilization success, a measure that is dependent on the ability of rival sperm competitors to fertilize the ova. It is well known that fertilization success may be conditional to genotypic interactions among males as well as between males and females. However, the consequences of effects arising from the random sampling of sperm competitors upon the estimation of genetic variance in sperm competitiveness have been overlooked. Here I perform simulations of mating trials performed in the context of sibling analysis to investigate whether the ability to detect additive genetic variance underlying the sperm competitiveness phenotype is hindered by the relative nature of fertilization success measurements. Results: Fertilization success values render biased sperm competitive ability values. Furthermore, asymmetries among males in the errors committed when estimating sperm competitive abilities are likely to exist as long as males exhibit variation in sperm competitiveness. Critically, random effects arising from the relative nature of fertilization success lead to an underestimation of underlying additive genetic variance in sperm competitive ability. Conclusion: The results show that, regardless of the existence of genotypic interactions affecting the output of sperm competition, fertilization success is not a perfect predictor of sperm competitive ability because of the stochasticity of the background used to obtain fertilization success measures. Random effects need to be considered in the debate over the maintenance of genetic variation in sperm competitiveness, and when testing good-genes and compatible-genes processes as explanations of polyandrous behaviour using repeatability/heritability data in sperm competitive ability. These findings support the notion that the genetic incompatibility hypothesis needs to be treated as an alternative hypothesis, rather than a null hypothesis, in studies that fail to detect intrinsic sire effects on the sperm competitiveness phenotype.

Sexual selection is thought to favor the evolution of secondary sexual traits in males that contribute to mating success. In species where females mate with more than one male, sexual selection also continues after copulation in the form of sperm competition and cryptic female choice. Theory suggests that sperm competition should favor traits such as testes size and sperm production that increase a male’s competitive fertilization success. Studies of experimental evolution offer a powerful approach for assessing evolutionary responses to variation in sexual selection pressures. Here we removed sexual selection by enforcing monogamy on replicate lines of a naturally polygamous horned beetle, Onthophagus taurus, and monitoring male investment in their testes for 21 generations. Testes size decreased in monogamous lines relative to lines in which sexual selection was allowed to continue. Differences in testes size were dependent on selection history and not breeding regime. Males from polygamous lines also had a competitive fertilization advantage when in sperm competition with males from monogamous lines. Females from polygamous lines produced sons in better condition, and those from monogamous lines increased their sons condition by mating polygamously. Rather than being costly for females, multiple mating appears to provide females with direct and/or indirect benefits. Neither body size nor horn size diverged between our monogamous and polygamous lines. Our data show that sperm competition does drive the evolution of testes size in onthophagine beetles, and provide general support for sperm competition theory.

Garcia-Gonzalez, F.2008.Male genetic quality and the inequality between paternity success and fertilization success: consequences for studies of sperm competition and the evolution of polyandry.Evolution, 62:1653-1665

Studies of postcopulatory sexual selection typically estimate a male’s fertilization success from his paternity success (P2) calculated at hatching or birth. However, P2 may be affected by differential embryo viability, thereby confounding estimations of true fertilization success (F2). This study examines the effects of variation in the ability of males to influence embryo viability upon the inequality between P2 and F2. It also investigates the consequences of this inequality for testing the hypothesis that polyandrous females accrue viability benefits for their offspring through facilitation of sperm competition (good-sperm model). Simulations of competitive mating trials show that although relative measures of male reproductive success tend to underestimate the strength of underlying good-sperm processes, good-sperm processes can be seriously overestimated using P2 values if males influence the viability of the embryos they sire. This study cautions the interpretation of P2 values as a proxy for fertilization success or sperm competitiveness in studies of postcopulatory sexual selection, and highlights that the good-sperm hypothesis needs empirical support from studies able to identify and separate unequivocally the males’ ability to win fertilizations from their ability to influence the development of embryos.

Two hypotheses could explain the evolution of paternal care: caring males are more attractive to females and mate more often (sexual selection); males care when the benefits in terms of offspring survival exceed the costs (natural selection). To test these hypotheses we used Phyllomorpha laciniata: females can choose whether to lay eggs on plants or on conspecifics, and the extent to which males carry eggs varies between populations. Our results do not support the sexual selection hypothesis: females did not choose to mate with egg-carrying males in either natural populations or experimental contexts. We compared two populations that differ in the extent of male egg carrying and we show that in the population where male egg carrying was more prevalent, parasitism pressure was higher. Field experiments revealed that, in the population with high parasitism rate, egg mortality as a result of parasitoid attack was up to 10 times higher on plants than on conspecifics. Egg carrying is thus an effective strategy that protects eggs against parasitoids. We conclude that the main benefit derived by males from egg carrying is an increase in offspring survival, and that males are sensitive to interpopulation differences in egg mortality risks. Male care in this system has evolved despite intermediate levels of paternity certainty because the impact on offspring survival is high, and the costs in terms of loss of mating opportunities low. Thus, our findings support the natural selection hypothesis, although additional work on more populations is needed to verify this.

In nonresource based mating systems females are thought to derive indirect genetic benefits by mating with high-quality males. Such benefits can be due either to the intrinsic genetic quality of sires or to beneficial interactions between maternal and paternal haplotypes. Animals with external fertilization and no parental care offer unrivaled opportunities to address these hypotheses. With these systems, cross-classified breeding designs and in vitro fertilization can be used to disentangle sources of genetic and environmental variance in offspring fitness. Here, we employ these approaches in the Australian sea urchin Heliocidaris erythrogramma and explore how sire–dam identities influence fertilization rates, embryo viability (survival to hatching), and metamorphosis, as well as the interrelationships between these potential fitness traits. We show that fertilization is influenced by a combination of strong maternal effects and intrinsic male effects. Our subsequent analysis of embryo viability, however, revealed a highly significant interaction between parental genotypes, indicating that partial incompatibilities can severely limit offspring survival at this life-history stage. Importantly, we detected no significant relationship between fertilization rates and embryo viability. This finding suggests that fertilization rates should not be inferred from hatching rates, which is commonly practiced in species in which it is not possible to estimate fertilization at conception. Finally, we detected significant additive genetic variance due to sires in rates of juvenile metamorphosis, and a positive correlation between fertilization rates and metamorphosis. This latter finding indicates that the performance of a male’s ejaculate in noncompetitive IVF trials predicts heritable offspring traits, although the fitness implications of variance in rates of spontaneous juvenile metamorphosis have yet to be determined.

A growing number of studies are suggesting that females can improve the viability of their embryos by mating with multiple males. However, the reason why females should have low rates of embryo viability is puzzling. Here we conduct a quantitative genetic study of maternal effects on embryo viability in the field cricket Teleogryllus oceanicus. After controlling for female body size, we find significant additive genetic variance for ovary weight, a measure of fecundity, and egg hatching success, a measure of embryo viability. Moreover, we show a genetic trade-off between these traits that is predicted from life-history theory. High rates of embryo mortality in this highly fecund species might therefore be explained by selection favouring an optimum balance between fecundity and embryo viability that maximizes maternal fitness. Paternal effects on female fecundity and embryo viability are often seen as benefits driving the evolution of polyandrous behaviour. However, we raise the alternative possibility that paternal effects might shift females from their naturally selected optimum, and present some support for the notion that sexual conflict over a female’s optimal fecundity and embryo viability might generate antagonistic coevolution between the sexes.

Spermatozoa exhibit taxonomically widespread patterns of divergent morphological evolution. However, the adaptive significance of variation in sperm morphology remains unclear. In this study we examine the role of natural variation in sperm length on fertilization success in the dung beetle Onthophagus taurus. We conducted sperm competition trials between males that differed in the length of their sperm and determined the paternity of resulting offspring using amplified fragment length polymorphism (AFLP) markers. We also quantified variation in the size and shape of the female’s sperm storage organ to determine whether female morphology influenced the competitiveness of different sperm morphologies. We found that fertilization success was biased toward males with relatively shorter sperm, but that selection on sperm length was dependent on female tract morphology; selection was directional for reduced sperm length across most of the spermathecal size range, but stabilizing in females with the smallest spermathecae. Our data provide empirical support for the theory that sperm competition should favor the evolution of numerous tiny sperm. Moreover, because sperm length is both heritable and genetically correlated with condition, our results are consistent with a process by which females can accrue genetic benefits for their offspring from the incitement of sperm competition and/or cryptic female choice, as proposed by the “sexy sperm” and “good sperm” models for the evolution of polyandry.

Although females are expected to maximize their reproductive success with only one or a few matings, the females of many species mate with multiple partners. Experimental studies have found evidence for an increase in egg or embryo viability when females mate polyandrously. These studies have been interpreted in the context of genetic-benefit models that propose that multiple mating increases offspring viability because it allows females to select male genotypes that influence viability directly or because it allows females to avoid genetic incompatibility. However, no studies have examined directly the precise mechanisms by which parents influence embryo viability. Using a morphological marker that enabled us to determine paternity and survival of embryos sired by individual male crickets in both sperm competitive and -noncompetitive situations, we show that males inducing high embryo viability enhance the viability of embryos sired by inferior males. These results indicate that paternal effects and interacting phenotypes determine embryo viability. They show that a male’s reproductive success is modified by the interaction between indirect genetic effects of sperm competitors. Importantly, our findings show that the benefits accruing to offspring of multiply mated females need not be transmitted genetically.

Females typically mate with more than one male despite the costs incurred, thus questioning Bateman’s principle. A series of genetic benefits have been proposed to account for the evolution of polyandry, including the acquisition of viability genes for offspring. The ‘intrinsic male quality’ hypothesis suggests that polyandry increases the probability that females produce offspring sired by males that bestow high viability on their offspring. Heritable variation in viability is the basic requirement for the occurrence of this genetic benefit. By using a half-sib breeding design with a species of cricket in which polyandry is known to increase hatching success, we present clear experimental evidence that intrinsic male quality contributes to embryo viability. Despite recent support for the evolution of polyandry based on compatibility of genotypes between males and females, we show that hatching success is not determined by an interaction between paternal and maternal genotypes but rather that sons inherit paternal genes that influence the viability of eggs laid by their mates. Moreover, our data implicate a potential role for indirect genetic effects of male accessory gland products on embryo viability. Additive genetic contributions to embryo viability may be an important factor underlying the frequently observed benefits of polyandrous behaviour.

1. The evolution of parental care and intraspecific parasitism involve conflicts of interest between mothers and other potential care givers who contribute to enhance offspring survival. In the golden egg bug, Phyllomorpha laciniata Villers (Heteroptera: Coreidae), females lay eggs on conspecifics and on plants. The adaptive significance of egg carrying in this species has been the subject of some controversy, which can only be resolved by determining the genetic relationship between the eggs and the adult who carries them. This study examined whether male acceptance of true genetic offspring occurs with a higher frequency than that expected from random oviposition on conspecifics. 2. Paternity analysis, using Amplified Fragment Length Polymorphism (AFLP) markers, was conducted on eggs carried by males housed with field-mated females. 3. Out of the total number of eggs sired by males in the experimental groups, the proportion of eggs carried by males that were their true genetic offspring was 30.8%. 4. Monte Carlo methods show that the probability of a male accepting an egg that is his true genetic offspring is higher than expected if females dumped eggs on males at random. 5. These results suggest that paternal care plays an important role in the maintenance of male egg carrying in this species. In addition, the methodology developed may become useful in determining true genetic parents in other species in which neither the father nor the mother can be determined by observational methods.

Female promiscuity often results in the ejaculates of different males competing to fertilize a female's ova. Experimental studies in insects have shown how sperm competition can be a potent selective force acting on an array of male reproductive traits, including features of the ejaculate such as sperm numbers or sperm size. However, experimental analysis of the role of sperm quality in determining paternity in insects has been neglected, despite the fact that sperm quality has been shown to influence the outcome of sperm competition in vertebrates. A recent comparative analysis found that males of polyandrous insect species show a higher proportion of live sperm in their stores, suggesting that sperm competition has shaped the quality of insect sperm. Here we test the hypothesis that sperm viability influences paternity at the within-species level. We use the cricket Teleogryllus oceanicus to conduct sperm competition trials involving pre-screened males that differ in the viability of their sperm. We find that paternity success is determined by the proportion of live sperm in a male's ejaculate. Furthermore, we were able to predict the patterns of paternity observed on the basis of the males' relative representation of viable sperm in the female's sperm storage organ. Our findings provide the first experimental evidence for the theory that sperm competition selects for higher sperm quality in insects, and indicate that between-male variation in sperm quality needs to be considered in theoretical and experimental studies of insect sperm competition.

In theoretical and experimental approaches to the study of sperm competition, it is often assumed that ejaculates always contain enough sperm of good quality and that they are successfully transferred and used for fertilization. However, this view neglects the potential effects of infertility and sperm limitation. Permanent or temporal male infertility due to male sterility, insemination failures, or failures to fertilize the ova implies that some males do not achieve sperm representation in the female reproductive tract after mating. A review of the literature suggests that rates of nonsperm representation may be high; values for the proportion of infertile matings across 30 insect species vary between 0% and 63%, with the median being 22%. I simulated P2 (the proportion of offspring fathered by the second male to copulate with a female in a double-mating trial) distributions under a mechanism of random sperm mixing when sample sizes and rates of male infertility varied. The results show that nonsperm representation can be responsible for high intraspecific variance in sperm precedence patterns and that it can generate misleading interpretations about the mechanism of sperm competition. Nonsperm representation might be a common obstacle in the studies of sperm competition and postcopulatory female choice.

Garcia-Gonzalez, F. and Gomendio, M.2004.Adjustment of copula duration and ejaculate size according to the risk of sperm competition in the golden egg bug (Phyllomorpha laciniata).Behavioral Ecology, 15:23-30

Several hypotheses have been proposed to explain the adaptive significance of prolonged copulations in insects, which include mate guarding and sperm loading functions. We have explored the adaptive significance of the prolonged copulations in the golden egg bug (copulations up to 50 h) and the effect of an increased risk of sperm competition on ejaculate investment. Our data support predictions derived from sperm competition theory, which posits that males are expected to increase ejaculate expenditure in response to an increased risk of sperm competition. Results show a combined response by males that has not been previously described: males in the presence of rivals increase copulation duration and the rate of sperm transfer. No relationship was found between male or female size and copulation duration or ejaculate size. Golden egg bug males transfer sperm slowly and gradually throughout copulation; thus an increase in the amount of sperm transferred and the corresponding increase in the male’s numerical representation in the female’s storage organs could be particularly important in a system in which so few sperm are transferred and in which so few sperm are stored by females. In addition, copulation duration may not only serve to increase the total amount of sperm transferred, but it may also increase the chances that the female will lay an egg soon after copulation has ended. This could explain why males tend to accept eggs after copulation, since they could be maximizing the chances that such eggs are fathered by them, and in this way they would substantially increase the survival rates of their offspring because eggs laid on plants suffer high mortality rates.

Theoretical models predict how paternal effort should vary depending on confidence of paternity and on the trade-offs between present and future reproduction. In this study we examine patterns of sperm precedence in Phyllomorpha laciniata and how confidence of paternity influences the willingness of males to carry eggs. Female golden egg bugs show a flexible pattern of oviposition behavior, which results in some eggs being carried by adults (mainly males) and some being laid on plants, where mortality rates are very high. Adults are more vulnerable to predators when carrying eggs; thus, it has been suggested that males should only accept eggs if there are chances that at least some of the eggs will be their true genetic offspring. We determined the confidence of paternity for naturally occurring individuals and its variation with the time. Paternity of eggs fertilized by the last males to mate with females previously mated in the field has been determined using amplified fragment length polymorphisms (AFLPs). The exclusion probability was 98%, showing that AFLP markers are suitable for paternity assignment. Sperm mixing seems the most likely mechanism of sperm competition, because the last male to copulate with field females sires an average of 43% of the eggs laid during the next five days. More importantly, the proportion of eggs sired does not change significantly during that period. We argue that intermediate levels of paternity can select for paternal care in this system because: (1) benefits of care in terms of offspring survival are very high; (2) males have nothing to gain from decreasing their parental effort in a given reproductive event because sperm mixing makes it difficult for males to reach high paternity levels and males are left with no cues to assess paternity; (3) males cannot chose to care for their offspring exclusively because they can neither discriminate their own eggs, nor can they predict when their own eggs will be produced; and (4) males suffer no loss of further matings with other females when they carry eggs. Thus, our findings do not support the traditional view that paternal investment is expected to arise only in species where confidence of paternity is high. The results suggest that females maximize the chances that several males will accept eggs at different times by promoting a mechanism of sperm mixing that ensures that all males that have copulated with a female have some chance of fathering offspring, that this probability remains constant with time, and that males have no cues as to when their own offspring will be produced.

Phyllomorpha laciniata Vill. (Heteroptera, Coreidae) females lay eggs on the host plant and on the backs of conspecifics. Since egg survival is greater when eggs develop on the backs of conspecifics than when laid on plants, we predict that females should prefer to lay eggs on conspecifics. In addition, because conspecifics are a high-quality site that represents a limiting resource, females should experience oviposition stimulation upon an encounter with a conspecific. Our results reveal that, when both the host plant and conspecifics are available simultaneously, females lay eggs preferentially on conspecifics. The results also support the second prediction, since females housed with conspecifics lay more than twice the number of eggs than isolated females. Isolated females do not seem to retain eggs, suggesting that oviposition stimulation is the result of an acceleration of egg-maturation rates. Other studies have found oviposition stimulation by mating and have suggested that it is the result of male strategies to increase short-term male reproductive success at some cost to females. The evolutionary scenario of our model organism seems to be quite different since females benefit greatly from increasing egg laying when there are conspecifics, because the advantages in terms of offspring survival are likely to translate into substantial increases in female reproductive success.

In natural populations of golden egg bugs (Phyllomorpha laciniata), females lay eggs on plants where they develop unattended, or on conspecifics, where they remain firmly glued until the nymphs hatch and start an independent life. Mortality rates among eggs laid on plants are higher than among eggs carried by adults. Because females cannot lay eggs on themselves, in order to improve offspring survival, they have to lay eggs on other individuals. Two hypotheses have been proposed to explain egg carrying: (1) the mating pair intraspecific brood parasitism hypothesis suggests that females dump eggs on copulating pairs, and (2) the paternal care hypothesis suggests that the system is driven mainly by males accepting eggs to improve the survival rates of their own offspring. Our data from the field show that 77% of the eggs are carried by males, because more males than females carry eggs, and because males carry a greater number of eggs. In addition, we show that mating males carry more recently laid eggs than single males. These results support the view that egg carrying is performed predominantly by males and that eggs are laid on males by their current mating partner, probably between repeated copulations. Males are likely to accept eggs, despite intermediate levels of paternity, because they cannot discriminate in favour of their own eggs, because rejected eggs will face 97% mortality rates on plants, and because they do not suffer mating costs when they carry eggs. However, females carry 23% of the eggs, but no differences in egg carrying have been found between mating and single females, suggesting that this is not the result of egg dumping while females are copulating. Egg carrying by females could reflect low levels of intraspecific parasitism, which is likely to reflect the low rate of successful attempts by egg-laying females who try to oviposit on other conspecifics rather indiscriminately, in an effort to improve the survival of their offspring.

Recent attention has focused on the role that sperm competition may play in the evolution of sperm morphology. Theoretical analyses predict increased sperm size, decreased sperm size, and no change in sperm size in response to sperm competition, depending on the assumptions made concerning the life history and function of sperm. However, although there is good evidence that sperm morphology varies widely within and between species, the adaptive significance of this variation has not been examined. Here we document significant intraspecific variation in sperm length in the field cricket, Teleogryllus oceanicus. Sperm length did not influence the rate of migration of sperm from the spermatophore to the female’s spermatheca. We performed sperm competition trials in which we varied the numbers of sperm transferred by each of two males that differed in the length of sperm they produced. Neither sperm length nor the number of sperm transferred influenced paternity. The same results were obtained using two different methods for assigning paternity. The distribution of paternity across a female’s mates was highly variable, with frequently one, or more in the case of females mated to four males, principal sire. There were no mating order effects on paternity. These data show that sperm do not mix randomly in the female’s spermatheca. We discuss several alternative explanations for the patterns of paternity observed.

Pitnick, S. & Garcia-Gonzalez, F.2002.Harm to females increases with male body size in Drosophila melanogaster.Proceedings of the Royal Society of London B, 269: 1821-1828

Previous studies indicate that female Drosophila melanogaster are harmed by their mates through copulation. Here, we demonstrate that the harm that males inflict upon females increases with male size. Specifically, both the lifespan and egg-production rate of females decreased significantly as an increasing function of the body size of their mates. Consequently, females mating with larger males had lower lifetime fitness. The detrimental effect of male size on female longevity was not mediated by male effects on female fecundity, egg-production rate or female-remating behaviour. Similarly, the influence of male size on female lifetime fecundity was independent of the male-size effect on female longevity. There was no relationship between female size and female resistance to male harm. Thus, although increasing male body size is known to enhance male mating success, it has a detrimental effect on the direct fitness of their mates. Our results indicate that this harm is a pleiotropic effect of some other selected function and not an adaptation. To the extent that females prefer to mate with larger males, this choice is harmful, a pattern that is consistent with the theory of sexually antagonistic coevolution.